Search Results (246)

Background/Objectives: The major limitations of self-nanoemulsifying systems include complex processing and expensive instrumentation required for solidification approaches. In this study, smart poloxamer-based solidification strategies were used to develop and optimize febuxostat-loaded formulations. Methods: A self-nanoemulsifying drug delivery system (SNEDDS) component was selected based on solubility and emulsification tests. The influence of poloxamer molecular weight (low or high) and its concentration (2–10% w/w) on formulation performance was assessed through the design of experiments. Finally, in-vitro melting assessment and a comparative dissolution test were performed on the optimized SNEDDS formulation. Results: Imwitor 988 and Tween 20 were selected to prepare the formulations. Increasing the molecular weight and concentration of the poloxamer significantly increased the temperature and time required for the melting of the SNEDDS formulation. The optimized SNEDDS formulation comprised 3.98% w/w poloxamer 188, which melts at 36 °C within 111 s. In-vitro melting showed that the formulation completely converted to a liquid state upon exposure to body temperature. Finally, the optimized SNEDDS formulation exhibited superior dissolution efficiency (96.66 ± 0.28%) compared to raw febuxostat (72.09 ± 4.33%) and marketed tablets (82.23 ± 3.10%). Conclusions: The poloxamer-based approach successfully addressed the limitations associated with conventional solidification while maintaining superior dissolution performance. Therefore, it emerges as a promising alternative approach for enhancing the bioavailability of poorly water-soluble drugs. Full article

Background: Artificial tooth wear impacts prosthesis durability and function; understanding material–antagonist interactions guides clinical choices. Aim: This in-vitro study aimed to assess the wear behavior of isosit and nanohybrid composite resin artificial teeth when opposed to various antagonist materials using 3D volumetric wear analysis. Materials and Methods: Sixty specimens (n = 10 per group) were prepared from two artificial tooth materials and assigned to six antagonist combinations: isosit–isosit, isosit–nanohybrid composite, isosit–porcelain, nanohybrid composite–isosit, nanohybrid composite–nanohybrid composite, and nanohybrid composite–porcelain. Specimens were scanned before and after 600,000 chewing cycles using a structured-light 3D scanner. Volumetric wear was calculated by superimposing pre- and post-test scans. Data were analyzed using two-way ANOVA and Tukey’s HSD test (α = 0.05). Results: Porcelain antagonists produced the highest wear values (p < 0.05). No significant difference was found between isosit and nanohybrid antagonists (p > 0.05). Identical material pairings showed less wear, though differences were not statistically significant. Conclusions: Porcelain as an antagonist increased wear risk. Using identical materials bilaterally, such as isosit–isosit or nanohybrid–nanohybrid, may help reduce artificial tooth wear in removable prostheses. Full article

Background: Automated semi-solid extrusion (SSE) material deposition is a promising new technology for preparing personalized medicines for different patient groups and veterinary applications. The technology enables the preparation of custom-made oral elastic gel tablets of active pharmaceutical ingredient (API) by using a semi-solid polymeric printing ink. Methods: An automated SSE material deposition method was used for generating chewable gel tablets loaded with propranolol hydrochloride (-HCl) at three different API content levels (3.0 mg, 4.0 mg, 5.0 mg). The physical appearance, surface morphology, dimensions, mass and mass variation, process-derived solid-state changes, mechanical properties, and in-vitro drug release of the gel tablets were studied. Results: The inclusion of API (1% w/w) in the semi-solid CuraBlend^TM printing mixture decreased viscosity and increased fluidity, thus promoting the spreading of the mixture on the printed (material deposition) bed and the printing performance of the gel tablets. The printed gel tablets were elastic, soft, jelly-like, chewable preparations. The mechanical properties of the gel tablets were dependent on the printing ink composition (i.e., with or without propranolol HCl). The maximum load for the final deformation of the CuraBlend™-API (3.0 mg) gel tablets was very uniform, ranging from 73 N to 80 N. The in-vitro dissolution test showed that more than 85% of the drug load was released within 15–20 min, thus verifying the immediate-release behavior of these drug preparations. Conclusions: Automated SSE material deposition as a modified 3D printing method is a feasible technology for preparing customized oral chewable gel tablets of propranolol HCl. Full article

Background: TMPRSS2 plays an important role in the viral entry mechanisms of influenza viruses and coronaviruses. Therefore, TMPRSS2 seems to be a suitable antiviral drug target. To exclude possible side effects of TMPRSS2 truncation in an early stage of drug in-vitro testing, this study aims to analyze the impact of TMPRSS2 truncation via antisense peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) targeting immune cells, using the example of regulatory T cells (Treg). Methods: TMPRSS2 was truncated in human Tregs using a splice-modulating PPMO. Effects on Treg function were analyzed by evaluation of surface marker and transcription factor expression, cytokine secretion, and effector cell suppression capability. Results: PPMO treatment led to a slight concentration-dependent toxicity in Tregs. Tregs with truncated TMPRSS2 behave similarly to untreated and control PPMO-treated cells in the analyzed assays. Conclusions: Treg function is not altered after TMPRSS2 truncation and therefore, no unwanted side effects in regard of Tregs are expected when using TMPRSS2-truncating PPMO as an anti-viral drug. Full article

Background/Objectives: This study aimed to develop a dry powder inhalation (DPI) formulation of rivaroxaban (RVX) using a combination of bead milling (BM) and jet milling (JM) to enhance lung-targeted delivery for the effective treatment of pulmonary embolism while minimizing systemic exposure. Methods: A carrier-free DPI formulation of RVX was developed using sequential BM and JM, with L-leucine incorporated at various concentrations (1%, 5%, and 10%) as a force control agent. The formulations were characterized for particle morphology, size distribution, crystallinity, and thermal properties. The in-vitro aerodynamic performance was evaluated using a next-generation impactor, while ex-vivo studies assessed anticoagulant activity. Pharmacokinetic and tissue distribution studies were carried out in Sprague Dawley rats following intratracheal administration, and the effects of inhaled RVX were compared with those of oral administration. Results: The optimized BM-JM-5L formulation achieved a Dv50 of 2.58 ± 0.01 µm and a fine particle fraction of 72.10 ± 2.46%, indicating suitability for pulmonary delivery. The two-step milling effectively reduced particle size and enhanced dispersibility without altering RVX’s physicochemical properties. Ex-vivo anticoagulation tests confirmed maintained or improved activity. In-vivo studies showed that pulmonary administration (5 mg/kg) led to a 493-fold increase in lung drug concentration and 2.56-fold higher relative bioavailability vs. oral dosing, with minimal heart tissue accumulation, confirming targeted lung delivery. Conclusions: The two-step milled RVX DPI formulations, particularly BM-JM-5L with 5% leucine, demonstrated significant potential for pulmonary administration by achieving high local drug concentrations, rapid onset, and improved bioavailability at lower doses. These findings highlight the feasibility of RVX as a DPI formulation for pulmonary delivery in treating pulmonary embolism. Full article

In-vitro spermatogenesis holds great potential in addressing male infertility, yet one of the main challenges is separating round spermatids from other germ cells in spermatogonial stem cell cultures. STA-PUT, a method based on velocity sedimentation, has been extensively tested for this application. Though somewhat effective, it requires bulky, expensive equipment and significant time. In contrast, the method of inertial microfluidics offers a compact, cost-effective, and faster alternative. In this study, we designed, fabricated, and tested a microfluidic spiral channel for isolating round spermatids and purifying spermatogenic cells. A commercially available spiral device close to the calculated specifications was tested for rapid prototyping, achieving 79% purity for non-spermatid cells in a single pass, with ability to achieve higher purity through repeated passes. However, the commercial device’s narrow outlets caused clogging, prompting the fabrication of a custom polydimethylsiloxane device matching the calculated specifications. This custom device demonstrated significant improvements, achieving 86% purity in a single pass compared to STA-PUT’s 38%, and that without any clogging issues. Further purification could be attained by repeated passes, as shown in earlier studies. This work underscores the efficacy of inertial microfluidics for efficient, high-purity cell separation, with the potential to revolutionize workflows in in-vitro spermatogenesis research. Full article

Raw milk and traditional fermented foods such as artisanal cheese represent a natural source of lactic acid bacteria (LAB). They can produce antimicrobial compounds, such as bacteriocins and lactic acid, which may be exploited in dairy biopreservation. This study aimed to conduct a systematic review and meta-analysis to synthesize the inhibition diameter (ID) of LAB against L. monocytogenes, S. aureus, and Salmonella spp. Literature electronic searches were performed on PubMed, Scopus, and Web of Science, to identify articles that reported data on in-vitro antimicrobial activity by LAB isolated from dairy foods. A total of 1665 papers were retrieved, and 20 primary studies were selected according to the selection criteria, of which 397 observations were extracted. Random-effects meta-regression models were employed to describe the effects of LAB genus, pathogen concentration, susceptibility method, incubation time, inoculation volume, agar type and pH on the IDs for L. monocytogens, S. aureus, and Salmonella spp. L. monocytogens was the most susceptible pathogen (p < 0.05) to the LAB effects, followed by S. aureus and Salmonella spp. As a whole, LAB from the Lacticaseibacillus genus were the most effective (p < 0.05) in inhibiting L. monocytogens (21.49 ± 2.654 mm), followed by S. aureus (21.06 ± 2.056 mm). Salmonella spp. presented higher (p < 0.05) susceptibility to Lactobacillus genus (19.93 ± 2.456 mm). From the results, a general trend could be observed for the well-diffusion method to produce higher (p < 0.05) ID estimates than the spot and disk methods (30.73 ± 2.530 mm vs. 21.98 ± 1.309 mm vs. 13.39 ± 1.403 mm for L. monocytogenes; 22.37 ± 1.073 mm vs. 14.91 ± 2.312 mm vs. 20.30 ± 2.319 mm for Salmonella spp.), respectively. Among the tested moderators, the pathogen’s inoculum concentration, the in vitro susceptibility assay itself, incubation time and inoculation volume on agar are determinant parameters to be looked at when designing a robust and reproducible experimental plan. The in vitro results reinforced that LAB can be useful in controlling the development of pathogenic bacteria frequently found in the dairy industry. Full article

Background: Recently, the demand for esthetic restorations has grown dramatically and extended into the pediatric population. The prefabricated zirconia crowns (PZCs) and custom-made zirconia crowns (CZCs) are new esthetic options in pediatric dentistry. However, they are still inadequately tested for use in children. Aim: To determine the fracture resistance and failure mode of the PZC in comparison to the CZC. Materials and Methods: In this in-vitro study, thirty cobalt-chromium dies were fabricated by scanning the negative replica of a prefabricated lower first permanent molar zirconia crown. CZCs were designed and milled using two different zirconia brands: Ceramill Zolid-FX (FX) and the Highly-Translucent (HT) zirconia. Dies were randomly assigned to receive either a PZC or a CZC (n = 10 in each group). All crowns were cemented on their respective dies using glass ionomer cement. Following artificial aging, all specimens were loaded to failure. Fracture mode analysis was performed. One-way ANOVA and Bonferroni post-hoc test were used for multiple comparisons across the groups. The significant level was set at p ≤ 0.05. Results: HT zirconia had a significantly higher fracture load compared to other groups (p < 0.05). The mean fracture resistance values were: (3087 ± 385) N for HT zirconia, (2633 ± 300) N for PZCs, and (2483 ± 381) N for FX, with no statistically significant difference in fracture strengths between PZCs and FX. Conclusions: HT zirconia crowns showed the highest fracture resistance amongst all groups. The fracture loads of tested crowns exceeded the maximum posterior biteforce. When placed in permanent molars, PZC are expected to perform well under masticatory forces in children. Full article

Objectives: The development of novel long-acting injectables for local anesthetics is necessary to effectively manage the acute postoperative pain. The aim of this study was to prepare an injectable oil-based formulation of ropivacaine (ROP) prodrug (ropivacaine stearoxil, ROP-ST) and to investigate the pharmacokinetics and pharmacodynamics after injectable administration. Methods: A novel N-acyloxymethyl prodrug of ROP, i.e., ROP-ST, was synthesized and its physicochemical properties such as log P, solubility and stability characterized. A soybean oil-based depot of ROP-ST was prepared, and the in-vitro release of ROP-ST was evaluated using an “inverted-cup” method. Pharmacokinetic profiles and tissue retention properties were investigated after intramuscular administration of the formulation in rats. The analgesic efficacy was assessed via a von Frey monofilaments test by measuring the paw withdrawal thresholds. Results: The structure of ROP-ST was ascertained with clear ¹H NMR assignment and accurate mass-to-charge ratio. The high Log P value of ROP-ST (9.16) demonstrated extremely low aqueous solubility, but the prodrug is biolabile when in contact with plasma or liver esterase. Intramuscular injection of ROP-ST oil solution in rats provided a significantly higher mean residence time without a very clear plasma peak of ROP. In a postoperative pain model of rats, the injection of ROP-ST oil solution into the vicinity of the sciatic nerve in the right ankle effectively controlled the postoperative pain for at least 72 h. Conclusions: The injectable oil-based depot formulation of N-acyloxymethyl prodrug of ROP may provide a new opportunity of long-acting local analgesia for postoperative pain. Full article

Background/Objectives: Skin wrinkles result from a myriad of multifaceted processes involving intrinsic and extrinsic aging. To combat this effect, plant stem cells offer a renewable and eco-friendly source for various industries, including cosmeceuticals. Salvia miltiorrhiza (SM), which contains the bioactive compound Rosmarinic acid (RA) and has been proposed for its anti-wrinkle effect. Methods: In the present study, calli from SM were cultured and Quality by Design (QbD) was implemented to investigate the effect of different types and concentrations of elicitors; jasmonic acid (JA) and salicylic acid (SA). Both raised RA levels yet, jasmonic acid (50 µM) has resulted in the highest yield for RA, at 16 mg/g. A nanofiber patch was prepared and characterized in-vitro by the release percentage, drug content, swelling degree, scanning electron microscope, and surface roughness. Then, the anti-wrinkle effect of the patch was tested in a UV wrinkle-induced mouse model. Results: Interestingly, after treatment, there were visibly fewer wrinkles, and the skin was softer than in the untreated control group. This study suggests that the treatment exerted its effect through the Nrf2/Keap1 pathway, which plays a crucial role in cellular antioxidant protective processes. By activating this pathway through boosting Nrf2 and diminishing Keap1 cellular content, the nanofiber patch enhances the production of antioxidant enzymes, such as superoxide dismutase and glutathione peroxidase, enhancesglutathione, and reduces the skin lipid peroxidation, collectively indicating enhanced skin quality. Conclusions: In conclusion, this study highlights the importance of this formula as an anti-wrinkle treatment, and future clinical studies are recommended to further unveil the potential of this formula. Full article

Triptolide (TP) is a diterpenoid compound extracted from the traditional Chinese medicinal herb Tripterygium wilfordii. It has antitumor and anti-inflammatory effects and stimulates immunity. However, its serious side effects, especially reproductive toxicity, limit its clinical application. This study employed a testicular injury model established by intraperitoneally injecting TP (0.2 mg/kg) in C57BL/6J male mice (age = 7–8 weeks) for 14 days. The control and TP mice’s testicular tissues were subjected to transcriptome sequencing to assess potential testicular damage mechanisms. Based on the transcriptome sequencing results and relevant literature reports, further experiments were performed. In addition, to alleviate triptolide-induced testicular damage, we treated the mice with N-acetyl-L-cysteine (NAC). The acquired data revealed that compared with the control mice, the TP-treated mice’s testes indicated severe damage. Transcriptome sequencing identified differentially expressed genes that showed enrichment in cell differentiation, apoptotic process, cell cycle, glutathione (GSH) metabolism, and the p53 signaling pathway. Furthermore, TUNEL assays and Western blot analysis showed that in the TP mice’s testicular tissues, the spermatocytes had mitochondrial pathway apoptosis as well as abnormal mitochondrial morphology and structure. Triptolide induces oxidative stress in testicular tissue by enhancing pro-oxidative systems and inhibiting antioxidant systems. NAC reduced testicular damage and apoptosis by alleviating TP-induced oxidative stress. This study also employed a GC2 cell line for in-vitro analyses, and the results were consistent with the in vivo experiments. This study provides evidence for alleviating TP’s adverse effects on the male reproductive system for better clinical application. Full article

Throughout history to the present day, infectious diseases have been a persistent global threat, causing significant harm to public health and economic stability. To address these challenges, the development of novel antimicrobial drugs is crucial. Hydrazones have gained significant attention in the scientific literature as promising candidates for developing new antimicrobial drugs. Two new hydrazones (H3 and H4) incorporating moieties that are known to enhance antimicrobial activity were synthesized. Methods: Hydrazone derivatives were synthesized through a condensation reaction of substituted acetophenone and nitro phenyl hydrazine. The compounds were characterized by their melting points and spectral analyses, including FT-IR, ¹H NMR, ¹³C NMR, and 2D NMR. Their antibacterial effects on Escherichia coli and Staphylococcus aureus were assessed in-vitro using the agar diffusion and broth dilution methods. Results: In-vitro testing demonstrated the compounds’ good activity against the tested organisms, particularly Gram-positive bacteria. At a concentration of 50 mg/mL, H3 produced a zone of inhibition (19 mm) comparable to that of the standard ciprofloxacin (20 mm) at 0.05 mg/mL. Only H3 was able to kill both Staphylococcus aureus and Escherichia coli at a concentration of 50 mg/mL. In all cases, H3 was found to be the most effective with optimum bactericidal and bacteriostatic activity against staphylococcus aureus and Escherichia coli. Conclusions: All the synthesized compounds were proven to possess promising antibacterial activity in vitro against the tested organisms. Full article

Background: This study aimed to optimize the coating process of Omega-3 fatty acid (OM3-FA) mini soft capsules containing the active pharmaceutical ingredients (APIs) pitavastatin and ezetimibe using near-infrared (NIR) spectroscopy for in-process monitoring. Cardiovascular disease treatments benefit from combining OM3-FA with lipid-lowering agents, but formulating such combinations in mini soft capsules presents challenges in maintaining stability and mechanical integrity. Methods: The coating process was developed using a pan coater and real-time NIR monitoring to ensure uniformity and quality. NIR spectroscopy enabled precise control of coating thickness, ensuring consistent drug distribution across the capsule surface. Results: The optimized process minimized OM3-FA oxidation and preserved the mechanical integrity of the capsules, as confirmed by texture analysis and in-vitro dissolution testing. This integration of NIR spectroscopy as a process analytical technology (PAT) significantly improved coating quality control, resulting in a stable and effective combination therapy for pitavastatin and ezetimibe in a mini soft capsule form. Conclusion: This approach offers an efficient solution for enhancing patient adherence in cardiovascular disease management. The application of NIR spectroscopy for real-time monitoring highlights its broader significance in pharmaceutical manufacturing, where it can serve as a versatile tool for ensuring product quality and optimizing production efficiency in diverse formulation processes. By incorporating NIR-based PAT, manufacturers can not only achieve product-specific improvements but also establish a foundation for continuous manufacturing and automated quality assurance systems, ultimately contributing to a more streamlined and robust production environment. Full article

Introduction: Poor outcomes following IVF treatments are speculated to be due to the transfer of aneuploid embryos that cannot be identified based on morphological evaluation alone. This leads to patients requiring numerous embryo transfers and, consequently, a prolonged time interval before live birth. Embryo selection following preimplantation genetic testing for aneuploidy (PGT-A) with next-generation sequencing (NGS) has been suggested as an intervention to shorten time to pregnancy in women undergoing in vitro fertilisation (IVF). Past studies assessing the clinical efficacy of PGT-A in improving clinical outcomes have been conflicting and the associated clinical pregnancy rates and live birth rates following the transfer of a mosaic embryos have yet to be determined. None of the existing studies solely included women of advanced reproductive age (ARA). The pilot study and proposed RCT will determine if, compared to morphological evaluation alone, the use of PGT-A through NGS is a more clinically effective, safer, and more cost-effective way to provide IVF treatment in women of advanced reproductive age. Method and Analysis: The proposed pilot study will aim to randomise 100 patients within a single-centre study to evaluate recruitment, randomisation, and adherence to study protocol and allocated trail arms by participating patients. The results of the pilot study will enable us to determine the sample size for a larger study to establish the effectiveness of PGT-A in ARA women. Ethics and Dissemination: The study (Integrated Research Application System Number 236067) received approval from the Health Research Authority and Health and Care Research Wales (HCRW) and the East Midlands—Leicester South Research Ethics Committee (20/EM/0290). The results will be made available to patients, the funders, the Reproductive Medicine societies, and other researchers. Trial registration: ClinicalTrials.gov Identifier: NCT05009745, n. Full article

Background: Advancements in genomic technologies are rapidly evolving, with the potential to transform laboratory diagnostics by enabling high-throughput analysis of complex biological data, such as microbiome data. Large Language Models (LLMs) have shown significant promise in extracting actionable insights from vast datasets, but their application in generating microbiome findings reports with clinical interpretations and lifestyle recommendations has not been explored yet. Methods: This article introduces an innovative framework that utilizes LLMs to automate the generation of findings reports in the context of microbiome diagnostics. The proposed model integrates LLMs within an event-driven, workflow-based architecture, designed to enhance scalability and adaptability in clinical laboratory environments. Special focus is given to aligning the model with clinical standards and regulatory guidelines such as the In-Vitro Diagnostic Regulation (IVDR) and the guidelines published by the High-Level Expert Group on Artificial Intelligence (HLEG AI). The implementation of this model was demonstrated through a prototype called “MicroFlow”. Results: The implementation of MicroFlow indicates the viability of automating findings report generation using LLMs. Initial evaluation by laboratory expert users indicated that the integration of LLMs is promising, with the generated reports being plausible and useful, although further testing on real-world data is necessary to assess the model’s accuracy and reliability. Conclusions: This work presents a potential approach for using LLMs to support the generation of findings reports in microbiome diagnostics. While the initial results seem promising, further evaluation and refinement are needed to ensure the model’s effectiveness and adherence to clinical standards. Future efforts will focus on improvements based on feedback from laboratory experts and comprehensive testing on real patient data. Full article